Composition for improving membrane composition and functioning of cells

ABSTRACT

It has now been found that after administration to a diseased person or person that is at risk for developing such disease of a neutraceutical or pharmaceutical composition that comprises
         a) a lipid fraction comprising at least one of docosahexaneoic acid (DHA), docosapentaenoic acid (DPA) and eicosapentaenoic acid (EPA);   b) a protein fraction comprising proteinaceous material from non-human origin which provide at least cysteine and/or taurine; and   c) a mineral fraction comprising at least one of manganese and molybdene, the health of these persons improves. Membrane function of several types of mammalian cells improves, which allows efficient treatment of immune related disorders, such as allergy, autoimmune diseases, cancer, cognitive dysfunction and other diseases of the nervous system, neuropathies, such as diabetic neuropathies and neuropathic pains, neuronal damage during insulin resistance, and gut diseases and support of the development of gut and lung function during growth or recovery.

RELATED APPLICATIONS

This application is a continuation of PCT application no.PCT/NL2006/000659 designating the United States and filed Dec. 22, 2006;which claims the benefit of the filing date of European application no.EP 05077972.7, filed Dec. 23, 2005; each of which is hereby incorporatedherein by reference in its entirety for all purposes.

FIELD

The invention relates to nutraceutical and pharmaceutical compositions,more in particular compositions that can improve membrane compositionand functioning of cells and through this the general cell functioning.

BACKGROUND

Membranes in living cells are known to have many important functions.The plasma membrane serves as a barrier to extra-cellular components(e.g. chemical compounds, antigens and cells); membranes (intra cellularas well as the plasma membrane) regulate the transport of componentsfrom the exterior to the interior of cells or organelles and vice versa.Membranes form a matrix for many endogenous reactions, for example manyenzyme controlled reactions that are important for the biosynthesis,metabolism or catabolism of compounds. Membranes are also important forrecognition and modulation of signals from the outside world andtherefore contribute to a proper response of the organism. Interactionwith the outside world includes the interaction of host cells withforeign cells or parts thereof like antigens/allergens, interaction ofhost cells with signal molecules like insulin, chemokines, cytokines andhormones and interaction of host cells with each other, such as theinteractions between nerve cells, in particular those within the centralnerve system (brain and in the spinal chord).

In the art many papers reveal particular aspects of the way living cellsinteract with extra-cellular signals. Phosphorylation of target proteinsin the membrane, as may occur after binding of a receptor with an extracellular ligand molecule, appears important for type and magnitude ofsignal transduction. Equally important is the assembly of signallingmolecule complexes with specific intracellular or membrane-boundproteins. These proteins, e.g. regulatory or scaffold proteins, maycomprise one or more specific domains such as the SH2, the SH3, the PTB,and the PTZ and WW domain types (see G. Cesarini et al. (eds.): ModularProtein Domains. Wiley VCH Verlag, 2004). The reaction of the cell to anoutside signal also depends on the nature of the membrane. In particularthe presence of some phosphatidylinositol (PI)-related compounds appearto be important for the function effected by proteins that possesspleckstrin homology (PH) domains, e.g. protein kinase C (PKC).

The way that a living cell reacts to an extracellular signal can varywidely and includes specific activation of phospholipid metabolism,specific cell adhesion behavior, a change in GTP-ase activity,localization of protein kinase activity, e.g. that of PKC, expression ofspecific transcription factors, regulation of the sensitivity of insulinreceptor, and the spatial arrangement or activity of receptors and ofion channels.

Interaction between nerve cells (neurons) requires activation of ionchannels, including the Na⁺, K+ and Ca2⁺ channels. In addition theregulation of numbers of receptors, also in relation to the differenttypes of receptors, is important for proper cognitive, emotional andsensorimotoric (like e.g. hearing, smelling, feeling and tasting)functions. Such regulation is also mediated by neuregulin (Nrg-1) andpostsynaptic density protein (PSD-95), whose functions appear to bedependent on membrane composition. In the end the signal transductionwill result in the activation of a variety of specific genes, whichtogether regulate consciousness, behaviour and cognitive andintellectual capabilities.

Cell membranes comprise many types of chemical components, likephospholipids, cholesterol, glycolipids, sphingolipids, sulfatides,cerebrosides, gangliosides, proteins, including glycoproteins, peptides,ions, vitamins, and many other components apart from water. Typicallythese components are not evenly distributed over the membrane. Polar andnon-polar areas can be identified. In particular to the so-called“rafts,” the often small non-polar areas in a plasma membrane, specificfunctions have been allocated (Brown, D. A., London, E., (2000) J. Biol.Chem. 275:17221-17224).

Membrane composition is subject to many changes over time due tophysical and/or enzymatic processes. Membrane-like structures also forma large part of the Golgi apparatus and the endoplasmatic reticulum (ER)and their structure determines their potential function and properties.

Several methods have been described to influence the interaction betweennerve cells and to improve cognitive function.

US 2005/0009779 discloses a method to increase membrane fluidity and toimprove receptor function in this way. It was claimed that saturatedfatty acids and cholesterol stiffen the membrane and that thecombination of long chain polyunsaturated fatty acids together withcertain components that improve methionine metabolism can make cellmembranes function in a better way. The document is e.g. silent aboutthe importance of the presence of polar membrane constituents other thanphospholipids and about the role of food constituents like manganese,molybdenum, taurine, cysteine, sulfate, water and the nature of thecarbohydrate fraction in suitable nutritional products for improvementof membrane function.

US 2005/0203053 discloses that neurological function could be improvedby enhancing the biosynthesis of phospholipids in the brain. This wasachieved by consuming a uridine source, a source of choline andoptionally a fatty acid. It was claimed that synaptic transmission wasimproved; the number of neurites was increased as well as thesensitivity of the P2Y receptor. Similar effects for these foodcomponents have been disclosed in US 2005/0176676.

WO 2004/028529 discloses a specific phospholipid comprising arachidonicacid, optionally in combination with medium chain triglycerides or asource of docosahexaenoic acid for improving cognitive function.

WO 2005/051091 discloses a specific blend of glycerophospholipids incombination with sphingomyelin and/or cholesterol, which blend resemblesthat of human breast milk and is present as a fat globule for use in themanufacture of infant formulae. The blend is claimed to be beneficialfor the development of cognitive and vision functions of the foetus,infants and children.

Specific lipid blends have been proposed for treatment of diseases ormodifying functioning of an organism.

In EP 1279400 an isotonic lipid emulsion is disclosed which comprises60-95 wt % MCT and 5-40 wt % fish oil and which comprises no vegetableoils. This lipid blend is claimed to rapidly modify cell membranes oforgans and tissues, and to be useful in the treatment of a wide range ofdiseases.

EP 0311091 discloses a specific lipid blend for parenteral use, whichcomprises MCT, ω-3 and ω-6 fatty acids, a phospholipid fraction andvitamin E. Such lipid blend is claimed to be useful for patients thatsuffer from surgery, multiple traumata, burns, infections, lung failureand chronic inflammatory diseases. The product as disclosed does notcomprise intact proteins or peptides and is not suitable for completenutrition.

EP 0484266 discloses nutritional products that comprise DHA and EPA,from about 24 to about 82 mg phospholipids per litre product and anucleotide mixture, which is claimed to be useful for nourishing infantsand for the dietary management of hepatic cirrhosis and diarrhea.

EP 1426053 discloses the use of phospholipids, sphingomyelin, orgalactolipid in the preparation of a pharmaceutical composition forparenteral use in inhibiting tumour cell adhesion, inhibition ofadhesion of metastasis cells and/or inhibition of tumour metastasisgrowth. The proposed formulations are unsuitable for complete nutritionby enteral route. No indication is given in what way other foodconstituents may interfere with the claimed efficacy.

Many papers speculate about the usefulness of eicosapentaenoic acid(EPA) or docosahexaenoic acid (DHA) for treating inflammatory diseases.However, serious difficulties have risen in demonstrating clinicalefficacy of the administration of these ω-3 fatty acids. Up to thefiling date predominantly in the treatment of arthritis a statisticallysignificant improvement could be observed.

It is an object of the present invention to change the composition ofcell membranes (particularly rafts) in several types of cells in amanner that is convenient, in particular by administration of a nicelytasting enteral formulation.

SUMMARY

In an aspect, the invention relates to a lipid fraction comprising atleast one component from the group of docosahexaenoic acid (DHA),docosapentaenoic acid (DPA) and eicosapentaenoic acid (EPA), the lipidfraction further comprising at least one component selected from thegroup of glycolipids for the improvement of immune function, themodulation of interaction between host cells and antigens, allergens,chemokines, cytokines, other host cells and/or insulin or the modulationof the ratio of the number of Th1 and Th2 cells.

The lipid fraction may in particular be a nutritional, nutriceutical orpharmaceutical composition, or a constituent thereof.

Accordingly, in an aspect, the invention now provides a nutritional,nutriceutical or pharmaceutical composition comprising at least onecomponent selected from the group of docosahexaenoic acid (DHA),docosapentaenoic acid (DPA) and eicosapentaenoic acid (EPA), the lipidfraction further comprising at least one component selected from thegroup of glycolipids.

In an aspect, the invention now provides a nutraceutical orpharmaceutical composition comprising:

a) a lipid fraction comprising at least one of docosahexaenoic acid(DHA), docosapentaenoic acid (DPA) and eicosapentaenoic acid (EPA);

b) a protein fraction comprising proteinaceous material from non-humanorigin which provide at least cysteine and/or taurine; and optionally c)a mineral fraction comprising at least one of manganese and molybdenum.

A composition (for use) in accordance with the invention preferablycomprises from about 50 to about 120 kcal per 100 grams.

In an embodiment the composition (for use) in accordance with furthercomprises a nucleotide fraction, selected from the group consisting ofnucleobases like uracil, cytosine, adenine, guanidine, thymine, theirphosphorylated forms (nucleotides), in particular themono-phosphorylated forms such as uridine monophosphate, but alsoadenosine and guanosine and other nucleosides.

Optionally, the composition further comprises a digestible carbohydratefraction, preferably including a source of galactose and/or ribose.

Preferably the lipid fraction is from dairy origin, from eggs or fromthe endosperm of plant seeds or beans and it preferably comprises fatglobules having a size of 0.001-10 μm.

Specific embodiments of the compositions are shown in the detaileddescription and the examples.

Also provided is the use of a composition according to the invention forimproving the function of a mammalian cell, selected from the group ofnerve cells, immune cells, stem cells, bone marrow cells anderythrocytes, for improving memory function in a mammal, for stimulatingimmune function in a mammal, for preventing obesity in a mammal, inautoimmune diseases and the like.

Also part of the invention is the use of said composition for themanufacture of a medicament for the treatment of dementia, such asAlzheimer's disease, neuropathies, depressed immune function, allergy,anemia, obesities, diabetes, autoimmune diseases, arrhythmias, heartfailures, tumours, COPD, bronchitis, arthritis, hepatitis, chronicinflammation, dyslipidaemia and rheumatoid diseases.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 demonstrates the significant improvement of biosynthesis andtransport of acetylcholine when feeding the product according theinvention to a rat, which is injected with AB.

DETAILED DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS

It has now been found that specific food compositions (also callednutraceutical compositions) or pharmaceutical compositions can improvethe functioning of living cells.

The products according the invention comprise a lipid fraction thatcomprises one or more of docosahexaenoic acid (DHA), docosapentaenoicacid (DPA) and eicosapentaenoic acid (EPA). In particular, the productsfurther comprise one or more glycolipids for use as a nutritional,nutriceutical or pharmaceutical composition for the improvement ofimmune function, the modulation of interaction between host cells andantigens, allergens, chemokines, cytokines, other host cells and/orinsulin or the modulation of the ratio of the number of Th1 and Th2cells.

Preferably the product according the invention comprises further aprotein fraction, the latter providing cysteine and/or taurineequivalents and optionally a mineral fraction that comprises manganeseand/or molybdenum. Optionally also a digestible carbohydrate fractionand/or a nucleotide source are beneficially included.

The lipid fraction preferably provides more than 30% of the energy thatis provided by the digestible components in the formula using ascalculation factors 9 kcal per g lipid, 4 kcal per gram protein or gramdigestible carbohydrate and zero kcal for the other components in theproduct. Preferably the product comprises a lipid content of 32-80, morepreferably 33-60 and still more preferably 36-50 percent of the totalenergy contents.

The amount of lipid fraction can be determined by applying the methodsknown in the art for measuring fat content in the food matrix asapplicable. For example, fat content for general foods is determined byapplying AOAC® official method 983.23, while the Roese-Gottlieb method(AOAC® 932.06) is better applicable for products based on dried milk(Lehner, R., Estoppey, A., (1954) Mitt. Lebensmitteluntersuchung Hyg.54:183-185), and again other methods are used for measuring lipidscontent in infant formulae or clinical nutrition.

The amount of individual components can be determined by applyingmethods specifically designed for measuring that specific component orby fractionating the fat fraction isolated from the extraction of thechloroform-methanol fraction as given in the 983.23 method. When twoaccepted methods would differ in a statistically significant way (usingP=0.05), the method, which gives the highest value is taken. The lipidfraction may include triglycerides, diacyl glycerides,monoacyl-glycerides, phospholipids, lysolecithins, glycolipids, sterolsand other lipid-soluble components. For the purpose of this applicationthe lipid fraction (and not the carbohydrate fraction) is defined toinclude sulfatides, cerebrosides, ceramides, sphingolipids,sphingomyelin, gangliosides and glycolipids like the globosides,hematosides and lactosylceramides. Specific analytical methods tomeasure the amounts of these components in a food matrix are known inthe art. These amounts should therefore be added to the amounts asmeasured by the other methods for determining total fat, as far as theyare not included in those measurements.

The polar components of the composition appear to have emulatingproperties which might interfere with the emulating systems which areconventionally used in the manufacture of homogeneous liquid formulae,which further comprise the protein, digestible carbohydrate andoptionally the mineral fractions in amounts according the invention. Byusing milk fractions as source of these glycolipids and glycoproteinsthe liquid forms of the products that are suitable for complete enteralnutrition remain homogeneous.

The total lipid fraction comprises long chain polyunsaturated fattyacids (having a chain length of at least 18 and abbreviated as LCP) ofthe omega 3 series (ω-3), and in particular one or more of DHA, EPA andDPA. These fatty acids can be present as triglycerides, phospholipids,sphingolipids, glycolipids or other food grade forms. The amount of ω-3LCP must be more than 400 mg per serving for adults, providing a dose of400-3300 mg of these fatty acids per day. The concentration of the sumof DHA, EPA and DPA in the lipid fraction is 2-50, preferably 5-45, morepreferably 20-40% of the lipid fraction. The concentration of the sum ofEPA, DHA and DPA in a liquid product is minimally 0.2 mg to maximally 15mg per ml, and preferably 0.3-12, more preferably 0.4-11 mg/ml. Forinfants the amount of w3 LCP usually should be more than 30 mg perserving, thus providing a dose of 30-1000 mg of these fatty acids perday.

In a specific embodiment, a composition of the invention comprises per100 g:

a) 1000 to 2000 mg of DHA+DPA+EPA+glycolipides;

b) 30-80 mg cysteine+taurine

c) 0.3-2 mg manganese+molybdene

The lipid fraction typically comprises a relatively low amount of longchain polyunsaturated fatty acids of the omega 6 series compared to theamount of other components of the lipid fraction. The ratio ω-3/ω-6 willusually be in the range 0.3-5, preferably 0.4-4, more preferably 0.5-2.The amount of ω-6 LCP's will be less than 50, preferably 5-40, morepreferably 8-30 wt %, most preferably 8-20 wt % and in particular 8-15wt % of the fatty acids in the formula.

Preferably also saturated fatty acids are included. These include thosefatty acids having a chain length of 8 to 24 carbon atoms. Preferablythese fatty acids include those having 10, 12, 14 and 16 carbon atoms.The amount of saturated fatty acids, if present, usually should be 6-60,preferably 10-40, more preferably 12-36% by weight of the lipid phase.In particular the amount of C14:0+C16:0 usually should be 6-60,preferably 10-40, more preferably 12-36 wt % of the lipid phase. Mediumchain triglycerides can origin from so called MCT oil, or origin fromcoconut oils or other suitable sources. The amount of MCT's usuallyshould be 1-60, preferably 3-40 wt % of the lipid fraction. Myristic andpalmitic acid can be included as acids, salts thereof, and esters withalkyl groups or acyl groups as in glycerides, or as other suitableforms. Preferably they are included as triglycerides, such as those thatoccur in palm kernel oil or in structured triglycerides with oneadditional ω-3 LCP or as diacyl or monoacyl glycerides. The monoacyl anddiacyl glycerides that comprise the saturated fatty acids are preferred,because they appear to provide technological advantages, compared to thetriglycerides forms, when higher amounts of lipids are included.

Oleic acid (C18:1) is preferably present in the lipid phase. It shouldbe present in a content of 5-40 wt % of the fatty acids, preferably6-35, more preferably 8-34 wt %. The amount in a liquid formula will bein the range of 0.1-2, preferably 0.15-1.2, more preferably 0.2-1.0 gper 100 ml liquid product.

Optionally an important part of the lipid fraction will comprise complexlipids like glycolipids, like sphingolipids such as sphingosines (e.g.sphingomyelin), acid sphingoglycolipids, such as sulfatides andgangliosides, and globosides. In the total lipid fraction the amount ofmaterial other than triglycerides and phospholipids is preferably morethan 0.01 wt %, preferably 0.015-1 wt %, more preferably 0.02-0.5 wt %.The amount of glycolipids is preferably at least 0.1 wt. % of the lipidfraction, in particular 0.1-20 wt. %. The amount of sphingomyelinpreferably should be more than 130 micromolar, preferably 135-3000micromolar, more preferably 140-2000 micromolar and the amount of acidsphingoglycolipids preferably should be more than 0.003, preferably0.005-0.5, more preferably 0.008-0.4 wt % for best results. The amountof gangliosides should be more than 7, preferably 8-80, more preferably9-40, in particular 9-25 mg/l liquid product. Preferred gangliosides areGD3 and GM3.

These compounds can be found in fractions that can be isolated frommicroorganisms like yeasts, bacteria, and in fungi, algae plants andanimal material, like egg, nerve tissue, fibroblasts, and milk. It ispreferred to use food grade ingredients. In particular it is preferredto use fractions from milk, such as extracts that comprise high levelsof fat globules. These can be obtained from the manufacture ofbuttermilk and in particular in the whey that remains aftermanufacturing cheese from buttermilk. Suitable products that arecommercially available include “buttermilk powder” and “butter serum.”

Milk whey comprises a relatively high amount of small fat globules,especially when macroproteins such as β-lactoglobulin have been removed.The concentration of small fat globules can be increased by applyingfiltration techniques on skimmed products, which concentrate the lipidlayer on one side of the membrane and remove molecules like salts andlactose. This product is called “whey microfiltrate retentate.”Fractions that are enriched with acidic sphingoglycolipids can also beisolated by applying chromatographic methods known in the art such asion exchange.

Milks from many mammals are suitable for isolation of the activecomponents, however, milk from mares, sheep, goat and camels areparticularly suited. It is most preferred to use a lipid extractisolated from sheep milk.

The amount of lipids from sheep milk should be more than 5 wt %,preferably 8-60 wt %, more preferably 10-40 wt % of the lipid fraction.

For example the fractions from egg, soy, colostrum, bone marrow, brain,milk and in particular buttermilk which are rich in fat globules, and inparticular the membranes of these fat globules are of particularinterest for inclusion in the products according the invention. Theseare concentrated in the complex glycolipids, which also compriserelatively high concentrations of carbohydrates other than glucose andfructose. Such fractions can be isolated by defattening the rawingredients by methods known in the art, e.g. by hexane extraction orcentrifugation. Additional methods should be applied to either theresidual lipid—or aqueous fractions to isolate the more polar lipidfraction present as fat globule membranes. Suitable methods includeselected removal of compounds of molecular weight below 300 such asminerals and lactose or removal of selected proteins, e.g. by usingfiltration, absorption, chromatography, osmosis techniques orprecipitation.

In particular fat globules of smaller size, in particular those having adiameter less than 4.0 um, preferably less than 3.0 um, more preferably0.001-2.0, most preferably 0.01-1.8 μm comprise the components ofinterest. In those situations wherein milk fat is not desired for thepatient, e.g. due to a wrong fatty acid profile, it is preferred toremove at least part of the fat globules of larger size, in particularthose having a diameter larger than 4.0 micrometer.

The lipid fraction from the milk should therefore preferably comprisemore than 1%, more preferably 1.2-20 and most preferably 1.3-16 wt %phospholipids. In addition the lipid fraction should preferably comprise0.1-20%, more preferably 0.2-10 and most preferably 0.6-8 wt %glycoproteins and glycolipids, and preferably 0.2-10, more preferably0.3-9, most preferably 0.35-8 wt % cholesterol or cholesteryl esters.Preferably the lipid fraction that is isolated from milk comprises theincreased amounts of at least two of phospholipids, glycoproteins,glycolipids and cholesterol and more preferably all components inamounts higher than in milk fat.

The lipid composition in the final product preferably comprises 1-80,more preferably 2-50, most preferably 3-40 wt % of the milk fractionenriched in components other than triglycerides and in particularcomprising 50% more glycolipids and/or glycoproteins originating fromsmaller fat globule membranes of milk compared to the amount in thelipid fraction of the natural material.

Suitable methods for isolating fractions, which are enriched in fatglobules, from dairy products can be found in U.S. Pat. No. 6,824,809,which is hereby incorporated by referencing. Also U.S. Pat. No.6,391,362 discloses a suitable way to isolate a fat globule fractionfrom the lipid fraction of milk or cream. However, it is preferred toisolate the smaller sized fat globules therefrom, e.g. by applying microfiltration to the second aqueous phase. It is preferred not tophosphorylate the original components of the fat globule.

An additionally preferred method for isolating suitable smaller fatglobules from milk is microfiltrating the aqueous phase that resultsafter butter, cottage cheese, or quark manufacture. Especially the milkof sheep, goats and camels are suitable sources for isolating the fatglobule fractions according to the invention, even though theconstituents of their fat globules differ to a large extent from thosein the fat globule of human milk. The fat globule fractions, which havebeen isolated according the methods as described, appear to be free frompotentially interfering substances like prions, pathogens or viruses.

Though the fat globules or fractions thereof, that origin from theingredients, are highly effective, they will not necessarily remainintact during the preparation or shelf life of the final productaccording the invention. The application of a homogenization step duringprocessing, and the inclusion of the ingredients, which causeeffectiveness of the final product, such as the protein fraction orother lipid components, interacts with the original fat globules thatare present in the ingredients. Therefore typically the size of the fatglobules in the product will increase and more than 50% of the lipidfraction will be provided as fat globules larger than 2 micron and innon-liposome form. More than 90% of the volume fraction of the lipidswill be present in globules having a diameter larger than 0.025 um.

The lipid fraction of the final product comprises per 100 ml liquidproduct preferably more than 0.56 g long chain polyunsaturated fattyacids, more preferably 0.57-3.1, most preferably 0.57-2.8 and inparticular 0.57-1.4 gram.

In those situations that the concentration in semi-solid and solidproducts is not explicitly mentioned or can not be directly derived fromthe claimed daily dose per daily portion, they can finally be calculatedfrom the claimed concentrations of the active ingredients in liquidproducts by correcting to percentage dry mass, assuming a dry masscontent of 15 g per 100 ml liquid product, 35 g per 100 g semi-solidproduct and 90 g mass per 100 g solid product.

In malnourished persons it is advantageous to include 0.1-10 g,preferably 0.2-5 g cholesterol sources per daily dose. Suitable sourcesof cholesterol include cholesterol, salts and esters thereof. Suchmalnourished patients may be hospital patients, patients that sufferfrom protein-energy malnourishment and the elderly. Not included in thisgroup are those malnourished persons that applied bad feeding practicesfor longer periods, which means in particular that for several monthsrelatively large amounts of trans fatty acids and polyunsaturated fattyacids, in particular of vegetable oils, which had become oxidized, wereconsumed.

The mineral fraction comprises at least manganese or molybdenum,optionally magnesium and zinc can be added. Preferably manganese andmolybdenum are both included and more preferably also magnesium isincluded in effective amounts.

Inclusion of manganese in a diet appears important for improvingmembrane function of cells, in particular the membrane function of nervecells. Especially those persons that are malnourished or have aninherited or metabolic disorder have imparted metabolic capacity forproducing sphingomyelin and/or related compounds like sulfatides andglycosylated ceramides. In addition, during these disorders or duringrapid growth persons experience increased needs for biosynthesis ofthese compounds. Examples of such persons are patients that suffer fromneurological diseases, like Alzheimer's Disease, Parkinson, depression,ME, persons that suffer from systemic lupus erythomatosus (SLE),paralysis caused by nerve damage or spinal chord injury, but also younginfants and foetuses, which are in a phase of growth of neural tissue,and pregnant women.

The amount of manganese administered/consumed per day should be morethan 7 mg preferably 9-300 mg for a person weighing 80 kg. Theconcentration in a liquid product can for example be calculated bytaking the daily volume and calculating the ratio of daily dose anddaily volume. Assuming a volume of 2000 ml for e.g. a tube feeding givesa manganese concentration of at least 7/2000 ml=350 microgram per 100ml. We also assume a caloric density of 1.0 kcal or 4.2 kilojoule per mlfor tube feed. The concentration per 100 kcal is therefore minimal 0.35mg per 100 kcal, preferably 0.45-15 mg/100 kcal or >0.83, preferably0.107-3.6 mg/100 kJ.

For infants weighing 3 kg and consuming 400 ml this minimumconcentration is 0.065-mg/100 ml. For infant formulae having a energydensity of 0.67 kcal/ml the concentration will therefore be more thanabout 0.1 mg per 100 kcal.

It was found that the inclusion of manganese as mentioned above had apromoting effect on the growth of lactobacilli in the gut, and severalof the benefits associated with this effect follow from this beneficialeffect, including a decrease of luminal pH, less growth of pathogens inthe gut and an increase in the strength and/or capacity of the immunefunction, measured in terms of a decreased rate of infections and/orless diarrhea. Also a better stool consistency was observed, inparticular in frequency of the occurrence of constipation. Despiteincreased use of manganese by these intestinal bacteria a systemiceffect was observed on cell function, especially when the concentrationsas given were administered.

Additional molybdenum is required to allow proper functioning of thecofactors, which appear important for creating a proper composition ofthe membranes, e.g. their sulfatide content, and in particularguarantees a proper functioning of nerve cells. Further, inclusion of aproper amount of molybdenum delays brain shrinkage in part of theelderly during aging. Molybdenum is also required to create a properbarrier function, e.g. of the gut, during maturation and growth orduring inflammatory bowel disease and/or food allergy.

A proper amount of molybdenum means an amount of more than 2 μg per kgbody weight per day, in particular 3-40, more preferably 3.6-32μg/kgbw.d. This will typically result in a concentration of 2.0-40ug/100 kcal [or 0.48-9.5 ug/100 kJ], preferably 2.6-30 μg/100 kcal [or0.62-7.1 ug/100 kJ] for infant formulae, and in 15-400, preferably18-100, more preferably 24-40 μg/100 kcal [equals respectively 3.6-95,preferably 4.3-23.8, more preferably 5.7-5.7 microgram per 100 kJ] forproducts for adults

The amount of magnesium that is consumed per day should be 400-1000 mg,preferably 480-800 mg for a person weighing 80 kg. When a completenutrition is designed this results in a concentration of 7.5-40 mg,preferably 9-30 mg per 100 kcal, which equals 1.8-9.5, preferably2.1-7.1 mg/100 kJ. Magnesium appears important to allow sufficientbiosynthesis of glycolipids.

Additional zinc can be included for several purposes, e.g. in order tonormalize copper homeostasis. For this it should be provided in a ratioof 8-40, preferably 10-30 fold excess to copper. Normalisation of copperhomeostasis is important to increase the degree of saturation ofceruloplasmin with copper and to prevent excess oxidation of membranefractions, including APP or parts thereof.

It appears important to have a balanced mineral composition. Thisenhances the efficacy per patient and also increases applicability in alarger group of the patients. Balance in this respect means that all ofthe minerals as mentioned above should be included, while also therelative amounts as indicated are important for creating a properbalance.

Suitable sources of these minerals include their salts with organic orinorganic anions like sulphate, carbonate, bicarbonate, hydroxide,chloride and citrate. Oxides of manganese and molybdenum should not beused, especially not in infant formulae. Preferably some sulphate formsare used, in such a way that sulphate provides 1-400, preferably 2-250,more preferably 3-125-mg/100 g product. Molybdenum is suitably includedas molybdate salt

It appeared that the effect of manganese and magnesium could beantagonized when high amounts of calcium were included in the product orwhen high doses of calcium were used in the total daily diet, forexample when nutritional supplements were consumed next to the productaccording the invention. The amount of calcium to manganese in theproduct should therefore be 1-235:1, preferably 5-205:1, more preferably10-100:1.

The protein fraction that is included in the product must be digestiblefor more than 80 wt % and preferably 90-100%. The protein fractioncomprises intact proteins, peptides as may be obtained by hydrolyses ofintact proteins and by syntheses, derivatives of peptides comprisingmore than 80% by weight amino acids, the (naturally occurring) aminoacids itself, and taurine, as well as derivatives of said amino acidsand taurine, like salts, N-acylated or N-acetylated forms as well asesters. Betaine, dimethylglycine, sarcosine, nucleosides material andcholine also comprise nitrogen, but will not be calculated as beingprotein.

The protein is due to provide available cysteine and/or taurineequivalents. All proteins that typically are used in food manufactureprovide cysteine. If considering dairy as a source of protein, it shouldbe realised that casein is a relatively poor source of cysteine. It istherefore advantageous to use at least partially a whey protein, and inparticular alpha-lactalbumin or whey proteins that are enriched inalpha-lactalbumin, in order to increase cysteine levels in the product.The amount of beta lactoglobulin is preferably relatively low comparedto that of alpha lactalbumin in relation to the ratio as observed innatural milk.

When milk is used as protein source, it is preferred to use a proteinfraction, which is also rich in proteins or peptides, which have a highdegree of glycosylation, such as casein glyco macropeptide (CGMP). Theseproteins/peptides are defined to be those peptides in which more than10% of the amino acid residues have been glycosylated. The amount ofCGMP in the fraction of the milk that is used for inclusion in theprotein fraction is preferably above 20%, more preferably more than 35,most preferably more than 40 wt %.

Preferably the protein comprises a histidine concentration of 2.5,preferably 3.0-6, more preferably 3.3-6 g per 100 g protein. This meansthat when whey proteins are used to a substantial degree free histidineneeds to be included.

The amount of taurine should be more than 0.1 g, preferably 0.2-4, morepreferably 0.4-3 g per daily dose. Typically this dose results inconcentrations in liquid products of more than 5 mg, preferably 7-100,more preferably 9-60 mg per 100 g product. Also taurine salts aresuitable ingredients.

Next to the above-described lipid, mineral and protein fractions, thecompositions of the inventions preferably comprise other fractions, suchas a carbohydrate fraction and/or a nucleotide fraction. Further, alsoinclusion of a vitamin fraction and/or a methyl donor fraction ispreferred.

The carbohydrate fraction comprises a digestible carbohydrate fraction,which preferably includes a source of galactose and/or ribose. In orderto provide sufficient energy in a carbohydrate blend that has a lowglycemic index the amount of these available and digestible non-glucosesugars must be less than 50, preferably 10-40 wt % of the amount ofglucose that is available to the consumer. Suitable sources of galactoseor ribose are disaccharides like lactose or food grade qualities of thesynthetic monosaccharides. Though the ribose from nucleotidescontributes to the physiological effect of the product, it is notcalculated to contribute to the total ribose content in the document,due to the relatively low doses of nucleotides in the product.

The digestible carbohydrate fraction beneficially comprisesglucosamines, like mannosamine and galactosamine or N-acylated formsthereof, for example N-acetylated forms, in an amount of 0.1-10,preferably 0.2-6 wt % of the digestible carbohydrate fraction.

Part of the carbohydrates, in particular 2-50 wt % can beneficially beironic acids as have been described in application EP 5103247.2. Part ofthe oligosaccharides, which have been disclosed in WO 2005/027663 isalso effective for the uses as mentioned below, in particular when usedwith the claimed protein—and mineral fraction.

The carbohydrate fraction preferably also comprises a source ofinositol. Myo-inositol is the preferred form, though up to 50% isallowed to origin from phosphorylated forms such as phytic acidequivalents such as phytates. Phosphatidylinositol also appears to be asuitable alternative.

The amount of inositol equivalents that should be administered to aperson weighing 80 kg per day is 0-1500, preferably 320-1200, morepreferably 380-1100 and most preferably 420-1000 mg. When calculatingthe desired daily dose for infants weighing 4 kg this results in 21-50mg.

Other ingredients that are beneficially included in the formula includea nucleotide source. Nucleotide sources are those ingredients that afterconsumption result in a raise in tissue levels of xanthine, nucleotides,nucleosides and/or nucleobases in vivo. Suitable ingredients include thenucleobases like uridine, cytidine, adenine, guanidine, thymidine, theirphosphorylated forms, in particular the mono-phosphorylated forms suchas uridinemonophosphate, but also adenosine and guanosine, nucleosidesand nucleotides.

Though beneficial effects can be observed by administering a singlenucleotide source, in particular uridinemonophosphate (UMP), to thecomponents of the claimed composition, about equally good effects,without risk of undesired side effects, can be obtained by usingmixtures of nucleotides, nucleosides or nucleobases. Such mixtures arepreferentially extracted from yeasts or animal tissues.

When UMP is used as source the amount per daily dose for a personweighing 80 kg must be 0.08-3 g, preferably 0.1-2, more preferably0.15-0.9 g. The required dosages of the equivalents on weight base canbe calculated from the dose of UMP by taking equimolar amounts of thenucleobases and correcting for molecular weight, using the molecularweight of UMP, which is about 324 Dalton.

Uridine derivatives like UDP, which is readily formed from dietetic UMP,appears important for transport of glycoproteins and glycolipids withinthe cell and availability thereof in the cytosol and plasma membrane.

In particular in an food for a child or infant, more in particular in aninfant formula, it may be desired that the ratio of the uracil source(including free nucleobase, nucleoside (uridine), and nucleotides (suchas UMP, UDP and UTP) to the sum of other nucleobases (including freenucleobases, nucleosides and nucleotides) is relatively high, i.e. thatthe uracil source or sources provides more than 25 mol %, in particularmore than 36 mol %, preferably more than 42 mol %, in particular atleast 50 mol % of the total nucleobase sources (including nucleosidesand nucleotides).

In particular in an food for a child or infant, more in particular in aninfant formula, the molar ratio [CMP+UMP] to [AMP+GMP+TMP+IMP] ispreferably more than 1.45, in particular more than 1.6, more preferablyat least 1.8, in particular at least 2.0. In particular, the ratio maybe up to 200, more in particular up to 100.

In particular in an food for a child or infant, more in particular in aninfant formula, the molar ratio UMP to [AMP+GMP+TMP+IMP] preferably ismore than 0.36, in particular more than 0.42, more in particular atleast 0.5, even more in particular at least 0.6. The ratio usually is upto 200, in particular up to 100.

In view of presently existing food regulations for infant formula, in aninfant formulation, the UMP concentration may in particular be in therange of 0.1-0.42 mg per 100 kJ and/or the total amount of nucleotidesmay in particular be in the range of 0.1-1.2 mg/100 KJ.

For improved effectiveness, it is considered advantageous to include UMPat a concentration of 0.42-10 mg/100 kJ, in particular 0.5-10 mg/100 kJ.The total amount of nucleotides may in particular be 100 mg/100 kJ.

The vitamin fraction, if present, should preferably comprise vitamin B6in a food grade form, which is able to increase the levels of pyridoxalphosphate in vivo in liver and/or brain. This means that the vitamin B6doses per day is restricted to not more than 50 mg and preferably notmore than 25 mg vitamin B6 per day for a person weighing 80 kg.Effective doses are 2 mg-50 preferably 2.4-40 mg, more preferably 8-25mg vitamin B6 for such a person. The optimal dose for a person having adifferent body weight can be calculated by taking the proportional part.For example for a baby weighing 3 kg the optimal dose is 37-1875 ug.Preferred forms of vitamin B6 are pyridoxine, pyridoxal, 3-acylatedpyridoxal analogues or salts thereof as disclosed in U.S. Pat. No.6,586,414, U.S. Pat. No. 6,548,519 and U.S. Pat. No. 6,339,085., whichare included by reference.

The vitamin fraction further can comprise biotin, folate and vitaminB12.

Biotin should be included in an amount of more than 1.5 μg/kgbw.d andpreferably 4-50, more preferably 5-40 μg/kgbw.d. This typically resultsin concentrations of biotin in the product of more than 6, preferably7-60, more preferably 8-40 μg/100 kcal for adults and >3, preferably4-50, more preferably 4.6-30 μg/100 kcal for infants. Suitable sourcesof biotin are known in the art.

Foliates include folic acid, folinic acid, methylated, methenylated andformulated forms of foliates, their salts or esters, as well as theirderivatives with one or more glutamic acid, and all in either reduced oroxidized form. Calculated to amount of folic acid the amount should beat least 300, preferably 420-2000 μg, more preferably 520-1500 μg/dailydose for an adult weighing 80 kg. For infants the recommended amount offolate per kg body weight is somewhat higher than for adults. It shouldbe >10 μg folic acid per kgbw.d and preferably 30-140, more preferably45-120 μg/kgbw.d.

A preferred embodiment of the formula comprises vitamin B12. Inparticular the vitamin B12 is not cyanocobalamine, but instead a memberselected from hydroxy- or methyl cobalamins or one of thethiolatocobalamines as disclosed in WO 02/087593 or the extracts asdisclosed in U.S. Pat. No. 6,187,761.

The other vitamins, such as vitamin A, D, E, K, C, B1, B2, B3 andpantothenic acid and minerals and trace elements, such as Na, K, Ca, Fe,Cu, Se, I, F comply with general recommendations for nutritionalrequirements, especially in infant formulae.

Additionally, a methyl donor is preferably included in the productsaccording the invention. A methyl donor is capable of generating atleast one methyl group when absorbed in the body of a mammal. Examplesof suitable methyl donors include betaine, choline, serine,dimethylglycine and sarcosine. It is preferred to select betaine ordimethylglycine for effective results. The advantages of includingadditional methyl donors in the product become in particular apparentwhen the protein source comprises more than 20 wt % protein of vegetableorigin, such as proteins from cereals like wheat, oats, rice, corn orsoy, or seeds/tubers like peas, lupine, potato, or beans. The effectivedose in such products is more than 0.18 mmol per kg body weight perdaily dose, preferably 0.19-2, more preferably 0.2-1.2 millimole perkgbw.d. When, for standardisation, the weight of a human body is set at70 kg, this would mean that choline (or dimethylglycine, because havingabout the same molecular weight) should be present in a liquidcomposition with more than 1.3 g per 2 liter product. A liquidsupplement for use by an adult consumer in an amount of 400 ml per daywould comprise 1.3 g per 400 ml.

The choline resulting from phospholipids is not calculated to contributeto this amount of methyl donor.

The fortification with one or more methyl donors is especiallyadvantageous if the amount of methyl acceptor remains low in the productin comparison with the amount of methyl donor. In particular it isadvantageous to have more than 3 fold molar excess of methyl donorscompared to the molar amount of methyl acceptors, such as glycine,phosphatidylethanolamine and polyamines such as spermine and spermidine.

Furthermore, statins can be beneficially included in those products thatare used by patients suffering from dyslipidemias and in particularhypercholesterolaemia. Admixture would be useful for periods shorterthan 2 weeks in those cases that a rapid decrease of plasma cholesterollevels is envisaged. The compounds may increase the efficacy of thecompositions of the invention during this period. However, adisadvantage on the longer term of including compounds like simvastatin,lovastatin, rosuvastatin, pravastatin, fluvastatin and atorvastatin istheir capability to inhibit potently the action of hydroxymethylglutaryl-CoA reductase when administered in relatively small amounts.Therefore, if the product is consumed for longer periods in conjunctionwith statins, the product should comprise one or more of the group ofubiquinons, ubiquinols, menaquinons, coenzyme Q10, dolichol, dolicholesters, dolichol ethers, synthetic poly-isoprenoids having more than 8isoprenoid units, such isoprenoids with one or more a carboxylic acidgroups, hydroxy groups having retinoids, lycopene, and sterols inamounts to counteract the disadvantages of administering statins.Preferably dolichol or its analogues are included, wherein the dolicholanalogues are substantially a polar for at least 80% of the molecule.Maximally fifteen percent, but preferably less than 10% of the carbonatoms is allowed to be substituted with polar groups such as hydroxy,alkoxy in particular methoxy, carboxyl, sulfhydryl, substitutedsulfhydryl, amino, substituted amino, alkyl or acyl moieties.

These inclusions are required to maintain efficacy of the product as animprover of immune function e.g. in cancer patients, and in particularas a product that decreases sensitivity to metastasis formation. Incombination with statin therapy the products even appear advantageous ifused without the protein and mineral fraction of the product in claim 1under the condition that the above-mentioned components are included inan amount of at least 20 mg dolichol analogues, at least 4 mg retinoids,at least 4 mg ubiquinons, ubiquinols, menaquinols or menaquinons, and atleast 0.02 mg sterols per daily dose. Suitable sterols are in particularnatural sterols as occur in mammalian organisms.

Effects

The products according to the invention normalize membrane function ofnerve cells, especially when these membranes are deformed and/or have awrong composition. Though one may speculate about the cause of thesedefects, it seems that a dysfunction of one or more membrane-boundenzymes, e.g. an upregulation of sphingomyelinase, which decreasessphingomyelin content and increases ceramides levels, seems to beimportant to this pathology. Also exposure to high amounts of stressors,a genetic disposition and/or chronic malnourishment may contribute tomore rapid development of imparted nerve functioning.

Persons older than 50 years of age are particularly prone to developingsuch condition, by a combination of aging and non-optimal nourishment.Prevalence of high amounts of nerve cells having deformed membranes willresult in e.g. impairment of cognitive function, sensory awareness andinsufficient capability to control the motoric system (muscles). Nervecells that possess deformed membranes will start to synthesize increasedamounts of AB (Abeta) proteins, which induce amyloid deposits, formingplaques and/or neurofibrillary tangles. The deformed membrane will alsostart to become a host for several lipophilic compounds like severalpeptides that are soluble in this matrix such as the dimers of AB, inparticular of AB 42, which may result in release of apoptosis signalsand death of nerve cells.

In the end this may lead to shrinkage of the brain volume and inparticular of the volume of the white matter, especially in the veryold. Syndromes caused by these effects are generally known as dementiaor Alzheimer's disease, although specific forms of these diseases havebeen described.

Neuropathy is partially caused under conditions wherein an organism andin particular the neural tissue is exposed for a longer period of timeto increased levels of reducing equivalents, in particular ofhydrogenated nicotinamide dinucleotide (NADH). Such levels are increasedin e.g. persons suffering from insulin resistance, like diabetics,persons that suffer from ischaemia (e.g. after surgery or after beensubjected to other physiological trauma which imparts blood flow totissues) or persons that abuse ethanol.

One may discriminate between neuropathies that occur centrally (brainand spinal chord) and peripheral neuropathies. The compositionsaccording to the invention are effective in both types. Central insulinresistance may result in pathologies as Alzheimer's or Parkinson'sdiseases. Peripheral neuropathy may impart motoric and sensoricfunctions and/or result in pains as explained below. Such neuropathiesmay also include neural function in specific organs like the liver.Without wanting to be bound by theory it may be important that thecompositions according the invention decrease cytosolic sialidaseactivity and increase the rate of biosynthesis of essential constituentsof nerve cells including sulfatides and gangliosides.

Neuropathy as caused by antibody attack also appears to be decreased.Without wanting to be bound by theory it seems important that theimprovement of the function of Schwann-type cells, in particular interms of repairing damaged myelin and cellular debris, contributes tothis effect, as does the improvement of the integrity of blood-nervebarrier and decrease of the activation state of glial cells in thespinal chord in such types of diseased persons. This makes thecompositions of the invention useful in the treatment of multiplesclerosis and in the decrease of chronic or neuropathic pain (contraryto normal pain).

Administration of the compositions according the invention appeared toimprove interaction between nerve cells, which effect had majorconsequences for many reactions of the body to outside triggers. Forexample proper notification and handling of sensory triggers andadequate definition of motoric functions as a reaction to these triggerswas improved. Interaction is essential to the thought process andawareness of emotions, including feelings of fear and pain (includingthreshold function).

In addition proper functioning of many organs and/or cells, that are atleast partially controlled by the nervus vagus or that react to localacetylcholine release e.g. by epithelial cells and macrophages, dependson the capability of nerve cells to handle these signals properly.Examples of such reactions include the activity of the gastrointestinaltract after sensory awareness of meal components, and the activity ofthe heart in rest (arrhythmias) and during exercise. Another example isthe activity of the immune system, in particular the immune cells thatcirculate in blood, in lymph and in particular in the epithelial cellsof lung and the gastrointestinal tract. These cells include the activityof cells that produce mucus (Goblet cells) and the glycocalyx, theantigen presenting cells, Paneth cells and Peyer's patches, andlymphocytes as locally present in the lamina propria, which are allstrongly influenced by their membrane composition and function.

The improvement of the functioning of the central nervous system,including the autonomous nerve system and the hypothalamus, includes thefunctioning of the white matter, and results in better thinkingcapabilities, enhanced emotional feelings, communication skills andmemory function.

It is claimed by the inventors that apart from neurotransmitter releaseand receptor function (notably G-protein receptors) also the compositionof the membranes of nerve cells is improved, and in particular theircontent of polar lipid-like components like sulfatides, ceramides,sphingolipids, glycoproteins and glycolipids in relation to the totalmass of the membrane and/or in relation to the amount of non-polarcomponents. Examples of non-polar compounds are cholesterol and dimericpeptides, in particular the amount of homo-dimeric non-polar peptides oflarger membrane proteins like the amyloid precursor protein, more inparticular the dimeric form of the AB-42 fragment. In this way it isaimed to change size and composition of membrane rafts and to maintain aproper functioning of the active parts of the rafts. In this way notonly the function of the receptor changes (which may be due to amodification of the affinity of ligand molecules for the receptors andan increase of discriminatory power, or a change in intrinsic activityof the ligands e.g. by influencing secondary messages such as the IP3pathway); also the activity of glycoproteins and membrane-bound enzymeschanges. In this way significant changes in metabolic and physiologicalactivities of living cells are achieved. This applies in particular tothe functioning of nerve cells, immune cells, bone marrow cells, stemcells and blood cells like erythrocytes. It was found by the inventorsthat acetylcholine biosynthesis and transport are improved byadministering the composition according the invention. The function ofmembrane-bound choline acetyltransferase (ChAT) was improved as well asits transport over the vesicle membrane into the synaptic cleft.Acetylcholine release is an essential part of brain function, whichincludes cognitive function and also of the activity of theparasympathetic nerve system, in particular the nervus vagus. This partof the nervous system regulates gastrointestinal behaviour and forexample heart rhythm. Improvement of heart arrhythmias, gut motility andgut-mediated systemic immune reaction is obtained by administering thecomposition according to the invention.

The compositions according the invention are also useful in repair ofdamaged nerve tissue. Nerve lesions can occur after trauma, for examplesurgery or spinal chord injury, but also in brain after chronic exposureto high diastolic blood pressure.

The compositions according to the invention are also useful in thetreatment of multiple sclerosis. The demyelination process is sloweddown as well as the creation of demyelinated plaques in the brain.

In patients suffering from Parkinson's disease, in particular idiopathicParkinson's disease (PD), the compositions according to the inventiondecrease the rate of deterioration of the substantia nigra cells. Inparticular the amount of amyloid plaques as well as the amount ofalpha-synuclein and Lewy bodies in brain will decrease in PD patients,but also in patients suffering from specific forms of dementias, e.g.Lewy body dementia, or suffering from multiple system atrophy (MSA) andamyotrophic lateral sclerosis (ALS).

Such precipitation of excess amounts of proteins in the brain, inparticular in neurons and glial cells, such as the occurrence ofpolyglutamate expansions during Huntington's disease, and tau-protein invarious forms of dementia, palsy and corticobasal degeneration, can beconveniently counteracted by intervention with the compositionsaccording the invention. Change of the intracellular membranes, e.g.those of the Golgi apparatus and/or endoplasmatic reticulum, and aconsequent different modification of the proteins may also play a rolein establishing this effect.

When used in administration to patients suffering from insulinresistance, and to babies or infants it is important to includecomponents, which increase insulin sensitivity, such as a proteinfraction comprising all essential amino acids and having a relativelyhigh aspartate to glutamate ratio, in particular above 0.42, preferably0.44-0.8, more preferably 0.45-0.6 or a vitamin fraction, whichcomprises at least riboflavin equivalents, vitamin B6 and/or biotin orpreferably all three vitamins. Examples of such protein fractionsinclude alpha-lactalbumin or protein blends to which aspartic acid hasbeen added.

In case the composition is administered to subjects suffering fromdiabetes, it is further beneficial to administer the composition asadjuvant to metformin and to refrain from co-administration with drugs,which are known to decrease insulin sensitivity. For subjects thatsuffer from insulin resistance it is further preferred to include afiber or fiber-like material that comprises resistant starch for morethan 20 wt % of the fiber blend in the composition. Also for diabeticsand obese subjects inclusion of a fiber-like material, such as slowlydigestible starch, galacto oligosaccharides, cellulose or fructooligofructose/oligofructose, is advantageous. for these groups a contentof about 30 wt % or more starch is advisable.

The active components in the compositions according to the inventionfurther regulate maturation of the eye and brain during fetaldevelopment. The compositions according the invention are thereforeuseful for administration to pregnant women, for allowing thedevelopment of good visual and brain functions in babies, includingcognitive and mental capabilities.

Improved communication between nervous cells appears to be important incombating epileptic convulsions, and reducing the frequency andintensity of schizophrenia, in decreasing the rate of attacks insubjects suffering from Gilles de la Tourette syndrome, in reducing ADHDand autistic behavior and in improving sleep behaviour.

For combating acute situations in these disorders, the compositionsaccording to the invention should be included in a ketogenic diet. Inparticular the caloric value of the nutritional product according theinvention must be more than 40% of the caloric value of the protein,lipid and digestible carbohydrate fraction in the product. Preferablythis amount is more than 46, more preferably more than 50%. In suchketogenic products the lipid fraction must meet the criteria asmentioned above. The amount of ω-6 long chain polyunsaturated fattyacids should be relatively low and the amount of saturated fatty acidsrelatively high.

The active components as present in the compositions according to theinvention also appear to be essential for proper maturation of the gutand proper development of its barrier, immune and transport function.This effect becomes evident from the reduced sensitivity to intestinalinfections, a reduced rate of diarrhea and bacterial overgrowth, andless sensitivity to food-born allergens of subjects that have obtainedsuch condition of the gut.

Without wanting to be bound by theory it is thought that it is importantthat the products according to the invention improve membrane functionof epithelial cells, of receptors like the CD1d receptor and that theyincrease the proper action of dendritic cells. This proper actionincludes the release of chemokines and cytokines, which recruit theright type of lymphocytes, provide a proper “Th1/Th2” response anddirect a proper migration of these cells to the lymph nodes. Theseeffects decrease the rate of pathogen translocation and pathogenrecognition. It is thought that the recruitment of the right types ofT-cells helps to combat undesired cells like allergens, pathogens andmutated cells and defines a proper response, e.g. an allergic reactionfor the organism. Due to the modified composition of the plasma membraneof e.g. endothelial cells, pathogens and/or allergens, in particularallergens that comprise a hydrophobic part, adhere less and/ordifferently, which causes a more appropriate reaction. In addition thecomposition of mucin produced by epithelial cells will improve, inparticular with regard to the amount of sulphated and sialylatedcomponents.

For obtaining maximal improvement of the condition of the gut,particularly in malnourished patients, the compositions according to theinvention should comprise a source of cholesterol. In particular theamount must be in the range of 2-100, preferably 6-60, most preferably9-40 mg per 100 g. Suitable sources are free cholesterol or cholesterylesters, in particular with oleic acid, or a mixture thereof, preferablyin a ratio of free to esterified cholesterol from 4-10:1.

It is also thought that the above mentioned effects with respect to theimmune system can also be held responsible for the improvement inresponse to a proinflammatory trigger and the decrease in magnitude ofan autoimmune reaction. This appears applicable in the treatment ofchronic inflammatory diseases, in particular arthritis, hepatitis,pancreatitis and chronic bronchitis and in the treatment of younginfants susceptible to developing diabetes type I and older adults whichappear to develop rheumatoid diseases, like rheumatoid arthritis andmultiple sclerosis.

The compositions according the invention facilitate lung development andproper repair of lung tissue, e.g. after burns, exposure to toxicants,emphysema, and chronic physical exposure like chronic coughing andduring inflammation and related lung infections and as results frompulmonary obstructive diseases like COPD and bronchitis.

Lung tissue comprises large amounts of specific surfactant molecules,whose rate of biosynthesis is increased by the compositions according tothe invention. The compositions are particularly useful for pregnantwomen for improving fetal lung development, for infants having smallgestational age, and in particular those that are sensitive to therespiratory distress syndrome. Problems with lung development can beidentified by analyzing at a proper moment, e.g. after 6 monthsgestation, the composition of the amniotic fluid and in particular theamount of sphingomyelin in relation to that of other lipid components.

When provided to patients for improving lung function the lipid fractionshould comprise a relatively high amount of palmitic acid (C16:0), inparticular more than 10 wt % of the fatty acids of the total lipidfraction, and more in particular 10.5-18-wt %. Palmitic acid shouldpreferably have for more than 15% the sn2 position in the triglyceridesfraction.

Efficacy of the formulations according to the invention in the treatmentof COPD can be established by measuring parameters indicative ofrespiratory capacity, e.g. FEV 1.0, and/or measuring the expression ofdifferent types of muscarinic receptors and chemokines secretion bysputum cells, like disclosed in Profita M., et al. (Allergy, 2005,60:1361-1369)

The compositions according the invention improve recognition ofundesired cells in the body such as mutated or tumor cells and changethe pattern of cytokine production by macrophages that were stimulatedby antigens. They therefore reduce the risk of developing tumors anddecrease the rate of recurrence of tumors after resection or destructionby other means, such as radio- or chemotherapy, and decrease the rate ofmetastasis formation.

The composition and constitution of the cell membranes appears toinfluence transport of reducing equivalents within the cell, for examplein the development of sensitivity to insulin. The compositions accordingthe invention therefore diminish the tendency to develop obesity duringchildhood but also during adult life.

In combating obesity, the compositions appear to decrease the amount ofcirculating C reactive protein (CRP) and to decrease the release of CRPby the adipose tissue. Endogenous synthesis of resistin by adiposetissue is increased. In this way the incidence of cardiovascular diseasein persons suffering from the metabolic syndrome, and in particular thedevelopment of arteriosclerosis and high blood pressure in obesepersons, is decreased.

Endocrine function of specific organs and organelles, which include thefunction of pancreas, hypothalamus and the Golgi and ER organelles inliver and vesicle biosynthesis in and release of endocrine messengersfrom nerve cells, is improved. The compositions according to theinvention are claimed to support release of insulin by the pancreas,secretion of digestive enzymes in cystic fibrosis patients, appropriatereaction of hypophyse to signals that arrive at or are produced by otherparts of the body, and immune function, especially the clearing functionof Kupffer cells as well as the biosynthetic role of organs that produceacute phase proteins.

The compositions can be beneficially used in combination with severaltypes of drugs for the purposes as mentioned above. The use of theproduct in combination with large planar molecules like statins has beendiscussed above, in particular in relation to the treatment of cognitiveimpairment in the elderly, which suffer from dyslipidaemia. Typically arelatively large part of these patients have a relatively high amount ofplaques and tangles.

The compositions of the invention can also be advantageously used incombination with phosphodiesterase inhibitors, when aiming improvementof cognitive function, in particular when the drugs are used for periodsshorter than about one month and when the dose of the inhibitors in thisperiod is less than 10 mg, preferably less than 2 mg per day for anadult.

The products according the invention are also advantageously used incombination with, or as adjuvant to vaccine therapy of amyloidformation. The rapid decrease in the amount of amyloid in brain issupported by biosynthesis of better membranes which thus prevents thedeleterious effects of residual excess amyloid.

Typically it requires at least several days before the affectivity ofthe compositions manifests itself. Optimal results are obtained byapplying administration regimens in humans, which achieve a daily use ofthe product for longer than 1 week, in particular longer than 2 weeks.The administration protocol may include a loading phase wherein duringthe first week 2 or 3 daily dosages are administered. Thereafter thedaily dosages which have been disclosed in this document have to beapplied for optimal results.

EXAMPLES Example 1 Introduction

In Alzheimer patients one of the hallmarks is the presence ofbeta-amyloid (Aβ) plaques. Formation of AB plaques causes a reduction inproduction capacity of acetylcholine, a neurotransmitter involved inlearning and memory processes. The reduction in acetylcholine levelsresults in memory loss [Isacson, 2002 #766]. Injecting rats with asolution containing AB results in similar problems in learning andmemory [Nakamura, 2001 #621]. This model is well accepted to test theeffects of beneficial compounds on AB-induced memory loss. Thisexperiment is addressing the effects of AB infusion on acetylcholineproducing cells and the transporter for acetylcholine. Furthermore, adietary composition is used to prevent these effects.

Experimental Design

Four groups of rats received either an infusion into the lateralventricle with beta-amyloid or a saline solution. Five weeks prior tothe infusions the rats were fed one of two diets, A or B. The fourgroups are summarized in table 1. Table lists the dietary composition ofthe two diets. Diet A serves as a control diet. Diet B is enriched inDHA, EPA and phospholipids to improve membrane quality. Furthermore, itis enriched in UMP and choline to stimulate membrane synthesis. And itis enriched in B-vitamins and folic acid to relieve homocysteine levels.

TABLE 1 Groups of Rats Differing in Infusion Solutions and Diet GroupInfusion Diet 1 (sham) Saline A 2 Abeta A 3 (sham) Saline B 4 Abeta B

TABLE 2 Composition of the Diets Diet A Diet B g/100 g fat g/100 g fatFatty acids LA 31.1 30.0 ALA 1.3 2.9 EPA 0.0 0.0 DHA — 3.3 total ω-631.1 30.4 total ω-3 1.3 13.0 ω-6/ω-3 23.5 2.3 mg/100 g food mg/100 gfood Phospholipids Soya Lecithin — 500 PC — 130 PS — 20 g/100 g foodg/100 g food Choline — 0.95 UMP — 1.55 mg/100 g food mg/100 g foodvitamins A 400 400 D3 100 100 E 3 253 K3 0.005 0.005 B1 0.4 0.4 B2 0.30.3 B6 0.6 4.725 B12 0.005 0.00575 Vitamin C 0 200 Niacin 2 2Pantothenic acid 0.8 0.8 Choline 86.8 86.8 Folic acid 0.1 1.35 Biotin0.2 0.2 minerals Iron 3.5 3.5 Copper 0.4 0.4 Zinc 1.2 1.2 Manganese 5 5Iodide 0.015 0.015 Selenium 0.009 0.159 Cobalt 0 0 Chromium 0.029 0.029Nickel 0.007 0.007 Fluor 0.09 0.09 Tin 0.1 0.1 Vanadium 0.009 0.009

Results

Table 3 (and FIG. 1) lists the effects of Aβ infusion and dietaryintervention on production and transport potential of acetylcholine. Aβinfusions in diet A fed rats resulted in a reduction of acetylcholineproducing cells (ChAT) and in transport capabilities for acetylcholine(VAChT). Feeding diet B completely restored production and transportcapacity of acetylcholine to normal levels.

TABLE 3 Results of ChAT and VAChT Diet A Diet B Sham Abeta Sham AbetaChAT 407.3 ± 29.27 309.4 ± 96.76 395.6 ± 62.6 404.0 ± 102.0 VAChT 296.3± 38.4  228.9 ± 53.31 271.4 ± 65.3 274.8 ± 58.8 

DISCUSSION AND CONCLUSION

Lateral ventricle infusions of Aβ induce a reduction in acetylcholineproduction and in transport capacity of acetylcholine. These effectswere completely diminished by feeding a diet enriched in omega-3 fattyacids, phospholipids, B-vitamins and UMP/choline.

Example 2

Ready-to-use liquid product for improving cognitive function in personsabove 50 years of age providing per 100 ml:

Energy 50-120 kcal; Protein 1-10 g; lipids 1-5 g; digestiblecarbohydrates 4-20 g, and which comprises

a—DHA+DPA+EPA=1000-2000 mg

b—30-280 mg cysteine or taurine

c—100-1000 mg phospholipids

d—0.5-3 mg vitamin B6

e—50-500 ug folic acid

f—1-30 ug vitamin B12

g—0.07-2 mg Manganese

h—0.07-2 mg molybdenum

Example 3

Ready-to-drink liquid for Alzheimer patients that comprises per 100 ml:

Energy 100 kcal Protein 3.06 g (casein, whey 80/20) Carbohydrates 13.3 g(maltodextrins, sucrose) Fat 3.73 g (fish oil, phospholipids) Comprising0.96 g DHA and 0.24 g EPA Uridine monophosphate 0.5 g (disodium salt)Choline 0.32 g Vitamin E 32 mg alpha-tocopherol Vitamin C 64 mg Selenium48 μg Vitamin B6 0.8 mg Folic acid 0.32 mg Vitamin B12 2.4 μg Magnesium20 mg Zinc 1.2 mg Manganese 0.3 mg Molybdenum 10 μgAlso 0.12 g NA, 0.15 g K, 0.12 g Cl, 80 mg Ca, 70 mg P, 1.6 mg Fe, 13.3μg I, 0.18 mg Cu, 6.7 μg Cr, 0.1 mg F, 0.16 mg vit A, 0.15 mg B1, 0.16mg B2, 1.8 mg B3, 0.53 mg B5, 0.7 μg D, 4.0 μg biotin and 5.3 μg vitaminK are included.

Example 4

Powdered anti-allergic infant formulae, which provide after freshreconstitution of 15.4 g powder per 100 ml water:

Energy 64-80 kcal Protein 1.0-2.0 g based exclusively on free aminoacids fraction or their salts, and which comprises at least 2 wt %cysteine and 0.2 wt % taurine Lipids 2-4 g which provides 2 wt % DHAbased on the total sum of fatty acids ingredients include 5-40 wt %buttermilk fat and 0.5-8 wt % egg lipids including lecithin Nucleotide1-40 mg Mixture of UMP and yeast extract equiv. Manganese 65-1000 μgincluded as salt, in particular MnSO₄ Molybdenum 2.3-300 ug included assalt, in particular molybdate salt Choline 20-200 mg included as cholinesalt, in particular equiv. choline bitartrateIn this example the determination of the amount of the additionalcomponents that are included, such as Na, K, Cl, Ca, P, Fe, Cu, Zn, Se,Cr, I, vit A, D, E, K, B1, B2, B3, B5, B6, B12, B11, biotine and Cfollows recommendations for infant formulae. Also myo-inositol wasincluded.

Example 5 Product for Patients that Suffer from Depressed ImmuneFunction

Liquid ready-to-use product that comprises per 100 ml:

Energy 630 kJ (or 110 kcal) Protein 4.0 g (buttermilk whey/casein 20-80)Carbohydrates 12.4 (3 g lactose, maltodextrins, slowly digestiblestarch) Lipids 5.3 (1 g milk fat, 1 g marine oil, 0.6 g egg lipids, 2.7g vegetable oil) Manganese 1 mg Molydenum 0.1 mg Betaine 100 mg Fibre1.0 g (resistant starch, galactooligosaccharides 50/50)Na 0.1 g, K 0.2 g, C10.13 g, Ca 0.8, P 0.8, Mg 50 mg, Fe 2.4 mg, Zn 2.4mg, Cu 0.3 mg, F 0.15, Se 8.6 ug, Cr 10 ug, I 20 ug, vit A 0.12 mg RE, D1.1 ug, E 1.6 mg TE, K 8 ug, B1 0.23 mg, B2 0.24 mg, B3 2.7 mg NE,Pantothenic acid 1.0 mg, B6 0.8 mg, B180 ug, B12 0.4 ug, biotin 10 ug, C12 mg

Example 6 Product for Patients Suffering from Cancer, HIV, Bone MarrowTransplantation, Heart Failure or COPD

Liquid ready-to-use composition based on full fat pasteurized milk fromsheep or camels, which comprises per 100 ml:

0.4 g algae oil, 0.4 g Betaine, 0.4 g ribose, and 0.2 g histidine andfurther Na 50 mg, K 160 mg, C1100, Ca 150 mg, P 120 mg, Mg 52 mg, Fe 3mg, Zn 2.8 mg, Cu 0.4 mg, Mn 3 mg, F 0.2 mg, Mo 200 ug, Se 11 ug, Cr 13ug, I 25 μg, vitamin A 188 μg RE, D 1.3 μg, E 2.3 mg a-TE, K 10 μg, B10.3 mg, B2 0.3 mg, B3 3.4 mg NE, pantothenic acid 2 mg, B6 (aspyridoxal) 0.6 mg, folic acid 100 μg, B12 0.7 μg, biotin 7.5 μg, anddimethylglycine 80 mg.

Example 7 Product for Preventing Obesity

Liquid product that comprises per 100 ml

Energy 336 kJ Protein 4.0 g (3.6 g soy protein isolate, 0.2 g aspartate,0.2 g histidine) Lipids 4.0 g (dolichol 0.2 g, phospholipids 1 g, milkglobule membrane enriched fat fraction 0.6 g, marine oil 1.0 g, rapeseedoil 0.5 g, sunflower 0.7 g, corn oil 0.5 g) Carbohydrates 12 g (lactose2 g, maltodextrins 6 g, slowly digestible starch 2 g, 2 g sucrose)Manganese 1 mg Molydenum 0.1 mg Betaine 100 mg Fibre 1.0 g (resistantstarch, galacto-oligosaccharides 50/50)Na 0.1 g, K 0.2 g, C10.13 g, Ca 0.8, P 0.8, Mg 50 mg, Fe 2.4 mg, Zn 2.4mg, Cu 0.3 mg, F 0.15, Se 8.6 μg, Cr 10 μg, I 20 μg, vit A 0.12 mg RE, D1.1 μg, E 1.6 mg TE, K 8 μg, B1 0.23 mg, B2 0.24 mg, B3 2.7 mg NE,Pantothenic acid 1.0 mg, B6 0.8 mg, B11 80 μg, B12 0.4 μg, biotin 10 μg,C 12 mg

Example 8

Liquid ready-to-use product for diabetic patient suffering fromneuropathy that comprises per 100 ml:

Protein 4.75 g (Pea protein, buttermilk whey) Lipids 3.78 g (fatfraction from sheep milk comprising 20% fat globules from small size 0.4g, egg lipids 1 g, vegetable oils 1.98 g and 0.4 g marine oils)Carbohydrates 11.75 g Galactose 1.5 Palatinose 3.0 Slowly digestiblestarch 1.0 Fructose 0.2 Maltodextrins 3 Glucose 2 Isomaltooligosaccharides 1.05 Fibres 2.0 g Galacto oligosaccharides 1.0Resistant starch 0.6 Cellulose 0.1 Fructo oligofructose/oligofructose0.3 g Baker's yeast 0.3 g

Vitamins, Minerals According to Example 2 Example 9 Product for PregnantWomen

Protein 3.06 g (casein, whey 80/20) Carbohydrates 13.3 g (maltodextrins,sucrose) Fat 3.73 g (fish oil, glycolipids rich fraction from sheepmilk) comprising 0.96 g DHA and 0.24 g EPA Uridine monophosphate 0.5 g(disodium salt) Dimethyl glycine 0.6 g Vitamin E 12 mg alpha-tocopherolVitamin C 24 mg Selenium 38 μg Vitamin B6 0.8 mg Folic acid 0.32 mgVitamin B12 4.4 μg Magnesium 30 mg Zinc 1.5 mg Manganese 0.3 mgMolybdenum 20 μgAlso 0.12 g NA, 0.15 g K, 0.12 g Cl, 80 mg Ca, 70 mg P, 1.6 mg Fe, 13.3μg I, 0.18 mg Cu, 6.7 μg Cr, 0.1 mg F, 0.16 mg vit A, 0.15 mg B1, 0.16mg B2, 1.8 mg B3, 0.53 mg B5, 0.7 μg D, 4.0 μg biotin and 5.3 μg vitaminK are included. Dimethyl glycine galactose.

1. A lipid fraction comprising at least one component chosen from thegroup of docosahexaenoic acid (DHA), docosapentaenoic acid (DPA) andeicosapentaenoic acid (EPA) and at least one component selected from thegroup of glycolipids for the improvement of immune function, themodulation of interaction between host cells and antigens, allergens,chemokines, cytokines, other host cells and/or insulin, and/or for theinduction of a proper response of Th1 and Th2 cells, CD-1 receptor ordendritic cells.
 2. The lipid fraction according to claim 1,characterized in that the composition further comprises a proteinfraction, which provides cysteine and/or taurine and a mineral fractionwhich comprises at least one of manganese and molybdenum
 3. The lipidfraction according to claim 1 further comprising a nucleotide fraction,which comprises at least one compound selected from the group ofnucleobases like uridine, cytidine, adenine, guanidine, thymidine, theirphosphorylated forms, in particular the mono-phosphorylated forms suchas uridine monophosphate; nucleotides; and nucleosides, in particularadenosine and guanosine.
 4. The lipid fraction according to claim 1,wherein the lipid fraction comprises also an ω-6 long chainpolyunsaturated fatty acid (having a carbon chain length of 18 or more)in an amount of 8-30 wt % of the fatty acids.
 5. The lipid fractionaccording to claim, 2, wherein the lipid fraction comprises more than0.01 wt % of at least one component of the group of glycolipids.
 6. Thelipid fraction according to claim 1, wherein the glycolipid is chosenfrom the group of sphingolipids, such as sphingosines, for instancesphingomyelin; acid sphingoglycolipids, such as sulfatides andgangliosides; and globosides.
 7. The lipid fraction according to claim1, wherein the composition further comprises a digestible carbohydratefraction, preferably a source of galactose and/or ribose.
 8. The lipidfraction according to claim 1, wherein the lipid fraction is at leastpartly originating from a dairy product, from eggs or from the endospermof plant seeds or beans.
 9. The lipid fraction according to claim 1,wherein the lipid fraction comprises fat globules having a size of0.001-10 μm.
 10. The lipid fraction according to claim 1, wherein thelipid comprises 1.3-16 wt % phospholipids and/or 0.1-20 wt %glycoproteins and glycolipids.
 11. The lipid fraction according to claim1, characterized in that the composition comprises a protein fractionwhich has a weight ratio of aspartate to glutamate of at least 0.42. 12.A composition for improving cognitive function in elderly subjectshaving an energy content of 50-120 kcal, a protein content of 1-10 g, alipids content of lipids 1-5 g, and a content of digestiblecarbohydrates of 4-20 g, which comprises: a) 1000-2000 mg DHA+DPA+EPA b)30-280 mg cysteine or taurine c) 100-1000 mg phospholipids d) 0.5-3 mgvitamin B6 e) 50-500 ug folic acid f) 1-30 ug vitamin B12 g) 0.07-2 mgmanganese; and h) 0.07-2 mg molybdenum
 13. A powdered anti-allergicinfant formulation, which comprises after fresh reconstitution of 15.4 gpowder per 100 ml water an energy content of 64-80 kcal, and: a proteinfraction of 1.0-2.0 g, which is based exclusively on free amino acids ortheir salts, and which comprises at least 2 wt % cysteine and 0.2 wt %taurine; a lipids fraction of 2-4 g, which comprises 2 wt % DHA based onthe total sum of fatty acids, wherein the lipid ingredients include 5-40wt % buttermilk fat and 0.5-8 wt % egg lipids including lecithin; anucleotide fraction of 1-40 mg, which comprises a mixture of UMP andyeast extract; a mineral fraction of 76.3-1300 μg comprising 65-1000 μgmanganese included as salt, in particular MgSO₄, and 2.3-300 μgmolybdenum included as salt, in particular molybdate salt; and 20-200 mgcholine, included as choline salt, in particular choline bitartrate. 14.The use of a composition as defined in claim 1 for improving thefunction of a mammalian cell, selected from the group of nerve cells,immune cells, stem cells, bone marrow cells and erythrocytes.
 15. Theuse of a composition as defined in claim 1 for stimulating immunefunction in a mammal.
 16. The use of a composition as defined in claim 1for preventing obesity in a mammal.
 17. The use of a composition asdefined in claim 1 for the manufacture of a medicament for the treatmentof an indication selected from the group of neuropathies, depressedimmune function, obesities, diabetes, autoimmune diseases, arrhythmias,heart failures, and chronic inflammation and infections caused bydiabetes.